Validation of a next-generation sequencing-based protocol for 24-chromosome aneuploidy screening of blastocysts.

Objective: To validate a 24-chromosome aneuploidy preimplantation genetic screening protocol based on multiple annealing and looping-based amplification cycle (MALBAC) and next-generation sequencing (NGS).

Design: Single-nucleotide polymorphism (SNP) array and MALBAC-NGS analysis.

Setting: University-affiliated in vitro fertilization (IVF) center.

Patient(s): Fifteen women from whom 30 blastocysts were obtained for genotyping.

Intervention(s): Not applicable.

Main Outcome Measure(s): Chromosomal status comparison of results of array comparative genomic hybridization (aCGH), SNP array, and MALBAC-NGS for 24-chromosome aneuploidy screening.

Result(s): Trophectoderm biopsy samples from blastocysts were first analyzed using array comparative genomic hybridization (aCGH); the embryos with detected with chromosomal abnormalities were rebiopsied, and dissociated into two portions, and subjected to SNP array and MALBAC-NGS for 24-chromosome aneuploidy screening. All 30 samples were successfully genotyped by array CGH, SNP array, and MALBAC-NGS. All blastocysts were correctly identified as aneuploid, and there was a 100% concordance in terms of diagnosis provided between the three methods. In the 720 detected chromosomes, the concordance rate between MALBAC-NGS and array CGH was 99.31% (715 of 720), and the concordance rate between MALBAC-NGS and SNP array was 99.58% (717 of 720). When compared with aCGH, MALBAC-NGS specificity for aneuploidy call was 99.85% (674 of 675; 95% CI, 99.17-99.97) with a sensitivity of 91.11% (41 of 45; 95% CI, 79.27-96.49). When compared with SNP array, MALBAC-NGS specificity for aneuploidy call was 99.85% (676 of 677; 95% CI, 99.17-99.97) with a sensitivity of 95.35% (41 of 43; 95% CI, 85.54-98.72).

Conclusion(s): MALBAC-NGS provides concordant chromosomal results when compared with aCGH and SNP array in blastocysts with chromosomal abnormalities.